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. 1998 Aug;51(4):232–233. doi: 10.1136/mp.51.4.232

A PCR-SSP method for detecting the His63Asp mutation in the HFE gene associated with hereditary haemochromatosis.

D Smillie 1
PMCID: PMC395644  PMID: 9893753

Abstract

Hereditary haemochromatosis is an autosomal recessive disease in which there is defective regulation of iron absorption, causing gradual accumulation of excessive amounts of iron in certain organs. Recently, a candidate gene for hereditary haemochromatosis has been identified, located on the short arm of chromosome 6, telomeric to the major histocompatibility complex (MHC) and showing sequence homology to the human leucocyte antigen (HLA) class I genes. Two mutations have been found in this gene that are potential markers for haemochromatosis. The first, a cysteine to tyrosine substitution (Cys282Tyr) is strongly associated with the disease, whereas the second mutation, a histidine to aspartic acid substitution (His63Asp) shows a less obvious relation. To examine the importance of this second mutation in hereditary haemochromatosis it is important to study the links between this genotype and abnormalities of iron metabolism. A polymerase chain reaction method using sequence specific primers is described which might be useful for identifying those individuals carrying the mutation that encodes the His63Asp substitution, who might be at risk from a milder form of haemochromatosis.

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Selected References

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  1. Aguilar Martinez P., Jeanjean P., Masmejean C., Guillard A., Biron C., Rabesandratana H., Schved J. F. Simple and rapid detection of the newly described mutations in the HLA-H gene. Blood. 1997 Mar 1;89(5):1835–1836. [PubMed] [Google Scholar]
  2. Becker I., Becker K. F., Röhrl M. H., Minkus G., Schütze K., Höfler H. Single-cell mutation analysis of tumors from stained histologic slides. Lab Invest. 1996 Dec;75(6):801–807. [PubMed] [Google Scholar]
  3. Beltinger C. P., Klimek F., Debatin K. M. Whole genome amplification of single cells from clinical peripheral blood smears. Mol Pathol. 1997 Oct;50(5):272–275. doi: 10.1136/mp.50.5.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bennett J. M., Catovsky D., Daniel M. T., Flandrin G., Galton D. A., Gralnick H. R., Sultan C. The morphological classification of acute lymphoblastic leukaemia: concordance among observers and clinical correlations. Br J Haematol. 1981 Apr;47(4):553–561. doi: 10.1111/j.1365-2141.1981.tb02684.x. [DOI] [PubMed] [Google Scholar]
  5. Beutler E., Gelbart T., West C., Lee P., Adams M., Blackstone R., Pockros P., Kosty M., Venditti C. P., Phatak P. D. Mutation analysis in hereditary hemochromatosis. Blood Cells Mol Dis. 1996;22(2):187–194b. doi: 10.1006/bcmd.1996.0027. [DOI] [PubMed] [Google Scholar]
  6. Beutler E. Genetic irony beyond haemochromatosis: clinical effects of HLA-H mutations. Lancet. 1997 Feb 1;349(9048):296–297. doi: 10.1016/S0140-6736(97)22005-2. [DOI] [PubMed] [Google Scholar]
  7. Böhm M., Wieland I., Schütze K., Rübben H. Microbeam MOMeNT: non-contact laser microdissection of membrane-mounted native tissue. Am J Pathol. 1997 Jul;151(1):63–67. [PMC free article] [PubMed] [Google Scholar]
  8. Emmert-Buck M. R., Bonner R. F., Smith P. D., Chuaqui R. F., Zhuang Z., Goldstein S. R., Weiss R. A., Liotta L. A. Laser capture microdissection. Science. 1996 Nov 8;274(5289):998–1001. doi: 10.1126/science.274.5289.998. [DOI] [PubMed] [Google Scholar]
  9. Feder J. N., Gnirke A., Thomas W., Tsuchihashi Z., Ruddy D. A., Basava A., Dormishian F., Domingo R., Jr, Ellis M. C., Fullan A. A novel MHC class I-like gene is mutated in patients with hereditary haemochromatosis. Nat Genet. 1996 Aug;13(4):399–408. doi: 10.1038/ng0896-399. [DOI] [PubMed] [Google Scholar]
  10. Going J. J., Lamb R. F. Practical histological microdissection for PCR analysis. J Pathol. 1996 May;179(1):121–124. doi: 10.1002/(SICI)1096-9896(199605)179:1<121::AID-PATH536>3.0.CO;2-D. [DOI] [PubMed] [Google Scholar]
  11. Guttridge M. G., Burr C., Klouda P. T. Identification of HLA-B35, B53, B18, B5, B78, and B17 alleles by the polymerase chain reaction using sequence-specific primers (PCR-SSP). Tissue Antigens. 1994 Jul;44(1):43–46. doi: 10.1111/j.1399-0039.1994.tb02355.x. [DOI] [PubMed] [Google Scholar]
  12. Jazwinska E. C., Cullen L. M., Busfield F., Pyper W. R., Webb S. I., Powell L. W., Morris C. P., Walsh T. P. Haemochromatosis and HLA-H. Nat Genet. 1996 Nov;14(3):249–251. doi: 10.1038/ng1196-249. [DOI] [PubMed] [Google Scholar]
  13. Jouanolle A. M., Gandon G., Jézéquel P., Blayau M., Campion M. L., Yaouanq J., Mosser J., Fergelot P., Chauvel B., Bouric P. Haemochromatosis and HLA-H. Nat Genet. 1996 Nov;14(3):251–252. doi: 10.1038/ng1196-251. [DOI] [PubMed] [Google Scholar]
  14. Merryweather-Clarke A. T., Pointon J. J., Shearman J. D., Robson K. J. Global prevalence of putative haemochromatosis mutations. J Med Genet. 1997 Apr;34(4):275–278. doi: 10.1136/jmg.34.4.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Miller S. A., Dykes D. D., Polesky H. F. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988 Feb 11;16(3):1215–1215. doi: 10.1093/nar/16.3.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moskaluk C. A., Kern S. E. Microdissection and polymerase chain reaction amplification of genomic DNA from histological tissue sections. Am J Pathol. 1997 May;150(5):1547–1552. [PMC free article] [PubMed] [Google Scholar]
  17. Smillie D. A PCR-SSP method for detecting the Cys282Tyr mutation in the HFE gene associated with hereditary haemochromatosis. Mol Pathol. 1997 Oct;50(5):275–276. doi: 10.1136/mp.50.5.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Takeuchi T., Soejima H., Faed J. M., Yun K. Efficient large-scale screening for the hemochromatosis susceptibility gene mutation. Blood. 1997 Oct 1;90(7):2848–2849. [PubMed] [Google Scholar]
  19. Zhuang Z., Bertheau P., Emmert-Buck M. R., Liotta L. A., Gnarra J., Linehan W. M., Lubensky I. A. A microdissection technique for archival DNA analysis of specific cell populations in lesions < 1 mm in size. Am J Pathol. 1995 Mar;146(3):620–625. [PMC free article] [PubMed] [Google Scholar]
  20. Zhuang Z., Roth M. J., Emmert-Buck M. R., Lubensky I. A., Liotta L. A., Solomon D. Detection of the von Hippel-Lindau gene deletion in cytologic specimens using microdissection and the polymerase chain reaction. Acta Cytol. 1994 Sep-Oct;38(5):671–675. [PubMed] [Google Scholar]

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